Compartmented neuron cultures for directional infection by alpha herpesviruses.
Compartmented neuron cultures for directional infection by alpha herpesviruses.
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DOI：
10.1002/0471143030.cb2604s43
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发表时间：
2009-06
影响因子：
--
通讯作者：
Enquist, Lynn
中科院分区：
文献类型：
--
作者：
Curanovic, Dusica;Ch'ng, Toh Hean;Szpara, Moriah;Enquist, Lynn
隔室神经元培养允许实验人员为神经元轴突和它们产生的体细胞建立单独的流体环境。通过在三室聚四氟乙烯环中培养神经元，实现了细胞体和轴突的物理隔离。分离的神经节被包裹在室的一端，轴突生长被引导在防水硅脂屏障下进入一个单独的室。由于轴突和细胞体位于不同的室室，它们可以分别感染和检测。我们描述的组装和使用区隔神经元培养在体外研究定向感染神经元由α疱疹病毒。选择性地将病毒接种物应用于一个隔室，可确保神经元的其余部分不受输入接种物的污染。这允许定量的病毒传播，并明确解释免疫荧光和电子显微镜图像。
Compartmented neuronal cultures allow experimenters to establish separate fluid environments for neuronal axons and the soma from which they emanate. Physical isolation of cell bodies and axons is achieved by culturing neurons in tri-chambered Teflon rings. Dissociated ganglia are plated in one end compartment of the trichamber, and axonal growth is guided underneath watertight silicone grease barriers into a separate compartment. Since the axons and cell bodies are located in different compartments, they can be infected and assayed separately. We describe the assembly and use of compartmented neuronal cultures for in vitro study of directional infection of neurons by alpha herpesviruses. Selective application of viral inoculum to only one compartment ensures that the remainder of the neuron is not contaminated by input inoculum. This allows for quantification of viral spread, and unambiguous interpretation of immunofluorescence and electron microscopy images.